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Fully Automated On-Chip Imaging Flow Cytometry System with Disposable Contamination-Free Plastic Re-Cultivation Chip
Kanagawa Academy of Science and Technology, KSP East 310, 3-2-1 Sakado, Takatsu-Ku, Kawasaki 213-0012, Japan
Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-Surugadai, Chiyoda, Tokyo 101-0062, Japan
* Author to whom correspondence should be addressed.
Received: 31 March 2011; in revised form: 18 May 2011 / Accepted: 26 May 2011 / Published: 7 June 2011
Abstract: We have developed a novel imaging cytometry system using a poly(methyl methacrylate (PMMA)) based microfluidic chip. The system was contamination-free, because sample suspensions contacted only with a flammable PMMA chip and no other component of the system. The transparency and low-fluorescence of PMMA was suitable for microscopic imaging of cells flowing through microchannels on the chip. Sample particles flowing through microchannels on the chip were discriminated by an image-recognition unit with a high-speed camera in real time at the rate of 200 event/s, e.g., microparticles 2.5 μm and 3.0 μm in diameter were differentiated with an error rate of less than 2%. Desired cells were separated automatically from other cells by electrophoretic or dielectrophoretic force one by one with a separation efficiency of 90%. Cells in suspension with fluorescent dye were separated using the same kind of microfluidic chip. Sample of 5 μL with 1 × 106 particle/mL was processed within 40 min. Separated cells could be cultured on the microfluidic chip without contamination. The whole operation of sample handling was automated using 3D micropipetting system. These results showed that the novel imaging flow cytometry system is practically applicable for biological research and clinical diagnostics.
Keywords: imaging cytometry; on-chip cytometry; cell sorting using microgel electrophoresis
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Hayashi, M.; Hattori, A.; Kim, H.; Terazono, H.; Kaneko, T.; Yasuda, K. Fully Automated On-Chip Imaging Flow Cytometry System with Disposable Contamination-Free Plastic Re-Cultivation Chip. Int. J. Mol. Sci. 2011, 12, 3618-3634.
Hayashi M, Hattori A, Kim H, Terazono H, Kaneko T, Yasuda K. Fully Automated On-Chip Imaging Flow Cytometry System with Disposable Contamination-Free Plastic Re-Cultivation Chip. International Journal of Molecular Sciences. 2011; 12(6):3618-3634.
Hayashi, Masahito; Hattori, Akihiro; Kim, Hyonchol; Terazono, Hideyuki; Kaneko, Tomoyuki; Yasuda, Kenji. 2011. "Fully Automated On-Chip Imaging Flow Cytometry System with Disposable Contamination-Free Plastic Re-Cultivation Chip." Int. J. Mol. Sci. 12, no. 6: 3618-3634.